Inflammation is accompanied by a substantial shift in tissue metabolism. One of the major metabolic signatures of inflammation is hypoxia, which is recently appreciated to significantly influence inflammatory disease outcome. Early in the disease process, such inflammatory hypoxia results, in large extent, from the recruitment of oxygen demanding inflammatory cell types, particularly neutrophils. In recent years, we have focused on defining targets and molecular pathways set into motion by inflammatory hypoxia. Results from these studies have defined a series of novel signaling mechanisms in which hypoxia (both in vitro and in vivo) drives the metabolism of extracellular nucleotides toward the generation of large amounts of extracellular adenosine. Central to this pathway was the identification of hypoxia-inducible factor (HIF) as an important regulator of the enzymes necessary for Ado metabolism (esp. CD73), thus identifying HIF-regulated Ado production as an endogenous anti-inflammatory pathway. Ongoing studies have revealed that Ado regulates HIF through the active deneddylation of Cullins, a family of proteins critical for the recruitment of ubiquitin ligases. Based on these preliminary studies, we hypothesize that Ado generated early in the inflammatory response functions as a feed-forward anti- inflammatory mechanism through direct actions on mucosal HIF stabilization. Three specific aims are directed at testing this hypothesis: In Specific Aim 1, we will define the contribution of neutrophils and epithelia to Ado generation at sites of inflammation. Specific Aim 2 will extend preliminary data to elucidate mechanisms of Ado-mediated Cullin-2 de-neddylation. Specific Aim 3 will Probe the role of Ado to HIF-mediated protection. The overall aim of this proposal is to identify novel metabolic signaling by HIF and Ado within the mucosa during inflammatory hypoxia.